Lima Et Al-2015-International Endodontic Journal

doi:10.1111/iej.
12328
Biocompatibility of root filling pastes used in

primary teeth
nior2, M. S. Rizzo3, R. D. Moura4, M. S. Moura5, M. D. M.

C. C. B. Lima1, A. M. Conde Ju
5
5
Lima & L. F. A. D. Moura
1
Postgraduate Programme in Dentistry, Federal University of Piau (UFPI), Teresina; 2Department of Morphology, Federal
University of Piau (UFPI), Teresina; 3Department of Clinic and Veterinary Surgery, Federal University of Piau (UFPI),
ao Paulo (USP), S~
ao Paulo; and 5Department of Pathology
Teresina; 4Postgraduate Programme in Pathology, University of S~
and Dentistry Clinic, Federal University of Piau (UFPI), Teresina, Brazil
Abstract
nior AM, Rizzo MS, Moura RD,
Lima CCB, Conde Ju
Moura MS, Lima MDM, Moura LFAD. Biocompatibility
of root filling pastes used in primary teeth. International
Endodontic Journal, 48, 405416, 2015.
Aim To evaluate the biocompatibility of two pastes

designed to fill the root canals of primary teeth.
Methodology A study group of 54 mice received
subcutaneous tissue implants of polyethylene tubes
containing CTZ or calcium hydroxide paste or, as a
negative control, empty tubes. Biocompatibility was
evaluated on days 7, 21 and 63, yielding a total of nine
groups of six animals each. Following the experimental
intervals, the implant areas were removed and subjected to histologic processing. After the tissues were
stained with HE and Masson trichrome, two pathologists performed a histologic analysis of the samples in a
blinded manner. Collagen fibre formation, tissue
thickness and inflammatory cell infiltration were
analysed qualitatively. Quantitative morphometry was
performed for the thickness, perimeter length and tissue
Introduction
The complex morphology of root canal systems and
the root resorption of primary molars are uncontrollable
factors that may hinder proper chemicalmechanical
Correspondence: L
ucia de F
atima Almeida de Deus Moura,
Department of Pathology and Dentistry Clinic, Federal University of Piau, Rua Angelica, 1650, Bairro de Fatima,
CEP 64049-532, Teresina, Piau, Brazil (Tel.: +55 86 3233
3050; e-mail: mouraiso@uol.com.br).
2014 International Endodontic Journal. Published by John Wiley & Sons Ltd
area of the region in direct contact with the open tube.

with the Tukey post-test and KruskalWallis
analysis followed by Dunns post-test, with significance
established as P < 0.05, were used for data analysis.
Results At 7 days, all groups had severe acute
inflammatory infiltrates. Inflammation was reduced at
21 days in the CTZ paste group. Mild chronic inflammatory infiltrates were observed after 63 days in the
CTZ and Ca(OH)2 paste groups; these groups also
showed a significant decrease in collagen fibre density
(P < 0.05), which was not observed in the control
group. The average tissue thickness, perimeter length
and area in contact with the tube decreased during
the experimental periods in all groups.
Conclusion The CTZ and calcium hydroxide pastes
demonstrated biocompatibility with subcutaneous tissue in this experimental model.
ANOVA
Keywords: biocompatibility, calcium hydroxide,

primary teeth, root canal filling materials, subcutaneous tissue.
Received 26 August 2013; accepted 29 May 2014
canal disinfection in primary teeth with pulp necrosis

(Harini Priya et al. 2010, de Sousa et al. 2011).
These conditions may also impede the development
of permanent tooth follicles as well as the establishment of accurate working length (Molander &
Dahlen 2003, Barja-Fidalgo et al. 2011, Gondim
et al. 2012).
Most of the recommended techniques for filling root
canals of primary teeth propose the use of instrumentation and the chemo-mechanical preparation of the root
canal system (American Academy of Pediatric
International Endodontic Journal, 48, 405416, 2015
405
Materials biocompatibility Lima et al.
Dentistry 2012). The use of calcium hydroxide (Ca

(OH)2) paste predominates over other techniques for the
pulp therapy of primary teeth. As a root filling material,
Ca(OH)2 paste demonstrates antibacterial action, biocompatibility and tissue repair capacity (Hauman &
Love 2003b, Mohammadi & Dummer 2011).
Alternative pulp therapies that have been developed
include those that recommend restricting treatment
to the pulp chamber and methods that may promote
the disinfection or sterilization of root canals with
necrotic pulps, as in the use of antibiotic-containing
pastes (Cappiello 1964, Takushige et al. 2004, Pinky
et al. 2011, Trairatvorakul & Detsomboonrat 2012).
Cappiello (1964) developed a paste for filling root
canals in primary teeth. The paste comprises chloramphenicol, tetracycline and zinc oxide with eugenol as a
vehicle and is currently known as CTZ paste (powder
base provided by a pharmacy in proportions of 1 : 1 : 2
chloramphenicol/tetracycline/zinc oxide, respectively).
This method obviates the need for canal instrumentation, thereby facilitating its use in young children and
users of public health services (Cappiello 1964).
Successful endodontic treatment of teeth with pulp
necrosis requires reducing or eliminating infection
within the root canal system. The medicaments used
in the pulp therapy of primary teeth should possess
antimicrobial activity and be biocompatible, particularly given the proximity of contact in the furcation
and periapical regions (Silva et al. 2010, Barja-Fidalgo
et al. 2011).
Biocompatibility is the ability of a material that is
in contact with a tissue to promote an appropriate
biological response with minimal inflammatory reactions. Methods that deploy substances in the subcutaneous tissues of laboratory animals are commonly
used to evaluate the biocompatibility of endodontic
materials (Hauman & Love 2003a, Viola et al. 2012).
The purpose of this study was to evaluate the biocompatibility of two pastes for the root canal treatment of primary teeth (CTZ and calcium hydroxide
pastes). The null hypothesis was that there is no difference between the biocompatibility of CTZ and calcium hydroxide pastes in mouse subcutaneous tissue.
Materials and methods

Ethical approval
The procedures used in this study were in accordance
with a protocol reviewed and approved by the Ethics
Committee on Animal Experimentation, Federal
406
University of Piau (UFPI) (opinion No. 067/12) and

with the ethical guidelines for animal experimentation
advocated in the European Communities Council
Directive of 24 November 1986 (86/609/EEC) regarding recommendations for the care and use of laboratory animals.
This experimental trial began in November 2012
and was concluded in May 2013 upon euthanasia of
the last test animal.
Animal model
A total of 54 mice (Mus musculus) were used. The
animals were Swiss albino males from 6 to 8 weeks of
age that weighed between 15 and 30 g. Throughout
the experiment, the animals were housed in a vivarium with standard rations and access to water ad libitum. All animals were examined every 2 days and
monitored for local or systemic changes.
The animals were wormed with Basken suspension (14.5 mg pyrantel pamoate + 9.5 mg oxantel
pamoate kg 1), which was administered orally in a
single dose 7 days prior to the onset of the experimental procedural.
Root canal pastes

The biocompatibility of the CTZ and calcium hydroxide pastes was tested using an empty tube as a negative control. Histological analysis was performed on
the tissue samples for each material at three different
intervals: 7, 21 and 63 days. The tissue response
was analysed for nine groups with six animals per
group, which were distributed as follows: Group I
six animals received a polyethylene tube containing
CTZ paste for 7 days; Group II six animals received
a polyethylene tube containing CTZ paste for
21 days; Group III six animals received a polyethylene tube containing CTZ paste for 63 days; Group
IV six animals received a polyethylene tube containing calcium hydroxide paste for 7 days; Group V six
animals received a polyethylene tube containing
calcium hydroxide paste for 21 days; Group VI six
animals received a polyethylene tube containing calcium hydroxide paste for 63 days; Group VII six animals received an empty polyethylene tube for 7 days;
Group VIII six animals received an empty polyethylene tube for 21 days; and Group IX six animals
received an empty polyethylene tube for 63 days.
The micronized powder base for the CTZ paste was
provided in 250-mg capsules by a compounding
Lima et al. Materials biocompatibility
pharmacy (Teresina, PI, Brazil), and the paste comprised

chloramphenicol
62.5 mg,
tetracycline
62.5 mg and zinc oxide 125 mg with 0.1 mL of
eugenol vehicle (Biodin^
amica, Ibipor~a, PR, Brazil).
Calcium hydroxide (Ca(OH)2) powder (analytical
grade) (Biodin^
amica) was provided in 250-mg capsules and reconstituted in 0.2 mL of distilled water
(Isofarma, Eusebio, CE, Brazil).
To prepare the two pastes, the powders were dispensed onto sterile glass plates and mixed with their
respective vehicles by a metal spatula at the time of
use.
Experimental design for subcutaneous implantation

Animals were pre-medicated with Tramadol at a dose
of 2 mg kg 1 (Tramadol hydrochloride 50 mg mL 1,
Hipolabor; Sabar
a, MG, Brazil) by deep intramuscular injection with a 1-mL syringe. After 10 minutes,
anaesthesia was induced using a combination of
xylazine (10 mg kg 1) (xylazine 2% Xilazin ; Syntec,
Cotia, SP, Brazil) and ketamine (80 mg kg 1) (ketamine hydrochloride 50 mg mL 1, Clortamina; BioChimico, Itatiaia, RJ, Brazil) prepared in the same
syringe and administered by deep intramuscular injection. The back of the animal was shaved, and the surgical region was disinfected with a 1% solution of
chlorhexidine gluconate.
In preparation for the dorsal implantation in each
animal, polyethylene tubes that were 1 mm in internal diameter (peripheral catheter type scalp No. 27
Solidor; Shijiazhuang, China) were sterilized in ethylene oxide and closed at one end with an Allis forceps.
The tubes were heated, sliced into 1-cm segments and
filled with the material to be tested. After a 0.5-cm linear incision was made at the caudal lumbar dorsum,
the soft tissue was dissected, and a tube was inserted
into the tissue. The skin incision was closed with 5.0
nylon sutures (Shalon Sutures; S~
ao Luis de Montes
Belo, GO, Brazil).
For the first 24 h following implantation, the animals received tramadol analgesia (2 mg kg 1), which
was administered by deep intramuscular injection
every 6 h.
At the end of 7, 21 or 63 days, animals from each
group were pre-medicated with tramadol (2 mg kg 1)
by deep intramuscular injection and euthanized with
an anaesthetic combination of xylazine (10 mg kg 1)
and ketamine (80 mg kg 1) prepared in the same
syringe and administered by deep intramuscular injection. The implant area, which comprised polyethylene
tubing, subcutaneous tissue and skin, was removed

and fixed in buffered 4% paraformaldehyde for 24 h.
Histologic analysis
Following fixation, the polyethylene tubes were
removed from the fixative, dehydrated in increasing
concentrations of alcohol, cleared in xylene, embedded in paraffin and blocked. The blocks were cut
using a microtome (Leica RM 2245) to yield 5-lmthick longitudinal semi-serial sections. Tissues were
stained with haematoxylin and eosin (HE) and Masson trichrome (Putt 1972).
The sections were examined under a binocular light
microscope (Olympus, Tokyo, Japan) at 100, 200 and
4009 magnification and digitally photographed.
A histologic analysis was performed by two pathologists, who were blind to the experimental conditions,
at different times. Both examiners analysed the same
fields. The Masson trichrome and HE stains were used
to analyse the collagen fibre formation and thickness,
respectively, at 2009 magnification. The inflammatory
infiltrate was analysed at 4009 magnification with HE
stain. The intra-examiner agreement was based on two
histologic evaluations performed at different intervals
on 10% of the sections, which were arranged in order
from the smallest to the largest increase (j = 0.80 and
0.81). The interexaminer agreement (j = 0.79) indicated interexaminer reliability.
The tissues that contacted the open end of the
tubes were analysed and graded for the following histologic findings: 1. formation of collagen fibres, 2. tissue thickness and 3. inflammatory infiltrate. The
density of collagen fibre formation was graded as 0
(absent), 1 (mild collagen fibres arranged similarly
to normal connective tissue), 2 (moderate individual
collagen fibres were observed, but did not display linear features or a typical wavy microanatomy) and 3
(severe very dense, individual collagen fibres cannot
be distinguished) (Silva et al. 2009, Queiroz et al.
2011).
The tissue thickness was assessed by the extent of
fibroblast cell layers formed and the presence of
macrophages in the peripheral contact region of the
tube and classified as follows: 0 (normal no layer),
1 (slightly increased 1 to 3 layers), 2 (moderately
increased 4 to 10 layers) and 3 (intensely increased
more than 10 layers). The inflammatory infiltrate
was analysed qualitatively to assess the characteristics
and concentration of polymorphonuclear cells (PMN)
and mononuclear cells, which were classified as
407
participated in the analysis and interpretation of

data and wrote the article. RDM participated in the
analysis of data. MSM revised the article. MDML
participated in the study preparation and revised
the article. LFADM conceived the study, participated
in the study preparation and coordination, and
wrote the article. All of the authors read and
approved the final manuscript.
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415
7a
Moderately increased (Fig. 3a)

Mild to moderately increased (Fig. 3d)
Severely increased (Fig. 3g)
Mild collagen fibre density (Fig. 2d)

Mild collagen fibre density (Fig. 2g)
Severe; mononuclear and occasional neutrophils

present (Fig. 4c)
Mild; predominantly mononuclear with occasional
neutrophils (Fig. 4f)
Mild; predominantly mononuclear with occasional
neutrophils (Fig. 4i)
Moderate; predominantly mononuclear, with the

possible presence of neutrophils (Fig. 4e)
Moderate; with macrophages, neutrophils and
neovascularization (Fig. 4h)
Moderately increased (Fig. 3c)

Mild to moderately increased (Fig. 3f)
Slightly increased (Fig. 3i)
Collagen fibres admixed with extracellular matrix

without any typical linear or wavy forms; individual
collagen fibres could not be distinguished (Fig. 2c)
Moderate (Fig. 2f)
Mild to moderate (Fig. 2i)
63c
Severe; presence of macrophages, lymphocytes,

neutrophils and newly formed vessels (Fig. 4b)
Severely increased (Fig. 3b)

Slightly increased (Fig. 3e)
Moderately increased, marked presence of fibroblasts
(Fig. 3h)
Mild collagen fibre density (Fig. 2e)

Individual collagen fibres observed as in normal
connective tissue (Fig. 2h)
Individual collagen fibres and alternating extracellular

matrix without linear features or wavy fibres (Fig. 2b)
21b
Experimental period (days)
P: KruskalWallis.
There was no statistically significant difference between groups (P > 0.05).
There was no statistically significant difference between groups (P > 0.05).
c
There was a statistically significant difference between the groups in the collagen fibre formation during this period (P < 0.05).
Inflammatory infiltrate (Fig. 4aj)

Empty tube
Severe; presence of lymphocytes,
macrophages, neutrophils, and newly
formed vessels (Fig. 4a)
Calcium hydroxide paste Moderate; mixed macrophages and
neutrophils present (Fig. 4d)
CTZ paste
Severe; predominantly mononuclear,
with neutrophils present (Fig. 4g)
Tissue thickness (Fig. 3aj)

Empty tube
Calcium hydroxide paste
CTZ paste
Calcium hydroxide paste

CTZ paste
Formation of collagen fibres (Fig. 2aj)

Empty tube
Mild to moderate (Fig. 2a)
Material tested
Table 1 Qualitative histologic analysis of the collagen fibre formation, thickness and inflammatory cell infiltration of the connective tissue adjacent to the open ends of the
polyethylene tubing
409
Ca(OH)2 paste
Empty tube
CTZ paste
(a)
(b)
(c)
(d)
(e)
(f)
(g)
(h)
(i)
7th Day
21st Day
63rd Day
(j)
Moderate
Mild
Severe
da
y
ay
3r
d
1s
tu
be
-6
-2
tu
y
pt
Em
Em
pt
y
pt
Em
td
da
y
be
tu
be
-7
3r
d
)2
H
O
a(
C
th
da
y
ay
-6
-2
)2
H
a(
O
a(
C
td
1s
th
-7
)2
H
TZ
C
da
y
da
y
3r
d
-6
-2
TZ
C
TZ
-7
1s
th
td
ay
da
y
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
Figure 2 (ai) Photomicrographs stained with Masson trichrome showing the formation of collagen fibres in the connective tissue adjacent to the open ends of the polyethylene tubes in contact with the empty tube (a, d, g), calcium hydroxide paste (Ca
(OH)2) (b, e, h) and CTZ paste (c, f, i), at 7, 21 and 63 days, respectively (2009 magnification). (j) Groups for the qualitative
analysis of the collagen fibre formation. (1) Space occupied by the tube and (2) subcutaneous tissue.
Discussion
A great challenge facing Paediatric Dentistry is to
develop biomaterials that possess antimicrobial properties, in addition to being biocompatible and resorb-
410
able, for primary teeth with necrotic pulps (Silva et al.

2010, Barja-Fidalgo et al. 2011, Queiroz et al. 2011).
CTZ paste has great potential as a treatment option
for endodontically compromised primary teeth in
Empty tube
Ca(OH)2 paste
CTZ paste
(a)
(b)
(c)
(d)
(e)
(f)
(g)
(h)
(i)
7th Day
21st Day
63rd Day
(j)
Slighty increased
Moderately increased
Intensely increased
3r
d
td
-6
tu
be
y
tu
be
-2
1s
th
-7
da
y
ay
da
y
Em
pt
Em
pt
H
O
a(
C
)2
tu
be
-6
-2
)2
H
O
a(
Em
pt
1s
3r
d
td
da
y
ay
da
y
-7
H
O
a(
C
TZ
)2
-6
-2
TZ
C
th
3r
d
td
1s
th
-7
TZ
C
da
y
ay
da
y
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
Figure 3 (ai) Photomicrographs showing the HE staining for the tissue thickness adjacent to the open end of polyethylene tube
in contact with the empty tube (a, d, g), calcium hydroxide paste (Ca(OH)2) (b, e, h) and CTZ paste (c, f, i), at 7, 21 and 63 days,
respectively (2009 magnification). (j) Groups for the qualitative analysis of tissue thickness. (1) Space occupied by the tubing, (2)
subcutaneous tissue, (*) neovascularization, (f) fibroblasts, (l) lymphocytes, (m) macrophages and (n) neutrophils.
411
Empty tube
Ca(OH)2 paste
CTZ paste
(a)
(b)
(c)
(d)
(e)
(f)
(g)
(h)
(i)
7th Day
21st Day
63rd Day
Mild
(j)
Moderate
Intense
da
y
1s
3r
d
td
ay
da
y
th
tu
be
-6
-2
be
Em
pt
y
tu
y
pt
Em
Em
O
a(
C
pt
y
tu
)2
be
-6
-2
)2
H
O
a(
-7
3r
d
td
1s
th
-7
H
)2
O
da
y
ay
da
y
da
y
3r
d
C
a(
TZ
-6
-2
TZ
C
TZ
-7
1s
th
td
ay
da
y
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
Figure 4 (ai) Photomicrographs of HE-stained tissue showing inflammatory infiltrate in the connective tissue adjacent to the
open end of the polyethylene tube in contact with the empty tube (a, d, g), calcium hydroxide paste (Ca(OH)2) (b, e, h) and
CTZ paste (c, f, i), at 7, 21 and 63 days, respectively (4009 magnification). (j) Groups for the qualitative analysis of the inflammatory reaction. (1) Space occupied by the tubing, (2) subcutaneous tissue, (*) neovascularization, (f) fibroblasts, (l) lymphocytes, (m) macrophages and (n) neutrophils.
412
Table 2 Mean values with standard deviations for thickness (lm), perimeter (lm) and area (lm2) of tissue reaction
Thickness (lm)
CTZ
Ca(OH)2
Control
P
Perimeter (lm)
CTZ
Ca(OH)2
Control
P
Area (lm2)
CTZ
Ca(OH)2
Control
P
Day 7
M (SD)
Day 21
M (SD)
Day 63
M (SD)
135.9 (59.0)
84.7 (17.2)
108.3 (41.6)
0.091
121.4 (69.6)
92.0 (28.5)
87.1 (22.6)
0.389
88.8 (41.0)
76.4 (26.1)
70.4 (31.8)
0.634
0.376
0.552
0.121
1046.1 (252.3)
884.4 (253.8)
1079.6 (73.1)
0.261
950.0 (211.8)
894.3 (143.0)
880.4 (115.2)
0.738
941.2 (179.0)
916.7 (173.4)
763.8 (190.9)
0.218
0.656
0.958
0.004
53554.9 (16442.2)
32132.6 (11382.7)
49362.2 (9999.3)
0.027
47344.8 (27953.0)
32404.8 (9832.4)
32561.9 (12713.6)
0.308
33816.3 (13998.1)
24996.0 (1335.6)
23483.9 (13930.3)
0.396
0.261
0.473
0.008
P*
M = mean; SD = standard deviation.

P: ANOVA test, applied between the materials in same experimental period.
*
P: ANOVA test, applied for each material between the different experimental periods.
young children with behavioural problems. Another

application for CTZ paste is for users of public
health services because of the large patient loads in
such facilities and the ease with which general
practitioners can perform these procedures due to the
technical simplicity, excellent clinical outcomes and
cost-effectiveness (Cappiello 1964, Bruno et al. 2007).
There is a lack of scientific literature on the use of
CTZ paste in clinical settings. This study is part of a
macro-development project that encompasses research
on the performance and antimicrobial action of CTZ
paste and its components.
Biocompatibility is the ability of an applied material
or substance to trigger the intended response in the
host. That is, although a biocompatible material may
not be inert, the reaction caused by a test material
must not pose an unacceptable physiological hazard
when compared with other scientifically approved
materials (Wataha 2001, Peters 2013).
Three tests are recommended for evaluating the biocompatibility of endodontic materials: initial, secondary and application. The method used in this study
was a sub-type that is used to evaluate biological tissue responses around the test material and to compare
such results to biocompatible controls with established
scientific acceptability (Olsson et al. 1981, ISO 7405
1996, ISO 10993-6 2007a, ISO 10993-12 2007b,
Hauman & Love 2003a, Silva et al. 2009, Pereira
et al. 2012). Subcutaneous implants, which were
introduced by Torneck (1966), are used most commonly in dentistry.
Compatibility with living tissue in the periapical or

interradicular region is one of the most important
properties for materials used in pulp therapy of primary teeth due to the occurrence of root resorption
and the presence of anatomical variations that may
jeopardize periodontal and periapical tissues, which
may in turn endanger the development of permanent
teeth (Lacativa et al. 2012).
Polyethylene tube implants are used experimentally
to simulate the relationship between the tooth apex
and periapical tissues because the material within the
tubes diffuses in a restricted area (Olsson et al. 1981,
International Organization for Standardization 2007a,
Silva et al. 2009, Khashaba et al. 2011, Marques
et al. 2011, Mutoh & Tani-Ishii 2011).
In this study, the groups in which empty polyethylene tubes were implanted had higher scores in the
qualitative histologic analysis for fibroplasia and had
significant differences in the perimeter and area of the
tissue regions in direct contact with the open tube.
This result was expected because no materials were
introduced that could induce an acute inflammatory
response or reaction. However, other studies have
reported responses to empty tubes, albeit less intense
than those observed for the experimental parameters
analysed at the time (Silva et al. 2009, Queiroz et al.
2011).
Significant reductions were observed in the density
of collagen fibres and the inflammatory reaction when
testing the calcium hydroxide paste for different
experimental periods. Calcium hydroxide is widely
413
used in dentistry due to its biocompatible properties

and antimicrobial, anti-inflammatory and reparative
activities (Hauman & Love 2003a,b, Mohammadi &
Dummer 2011, de Sousa et al. 2011, Gondim et al.
2012). The biocompatibility results obtained in this
study for calcium hydroxide paste are consistent with
those reported by several authors (Holland et al.
2001, Semenoff et al. 2008, Silva et al. 2010, Andolfatto et al. 2012).
In the present study, the tubes containing CTZ
paste displayed progressive darkening over time,
which may have been caused by the tetracycline
component. The CTZ paste was associated with an
initial intense inflammatory response during the first
days of the experiment. At the end of the experiment,
a mild chronic inflammatory process was observed,
with a qualitative and quantitative decrease in the
density of collagen fibres and tissue thickness. This
finding suggests that CTZ was biocompatible and
showed therapeutic properties. In a previous study,
histopathologic analysis of pulpotomy performed in
dogs teeth treated with CTZ paste demonstrated the
occurrence of an initial intense inflammatory process
in the coronal pulp with reduced inflammation at the
end of the experiment (Bruno et al. 2007). However,
the literature lacks studies on the biocompatibility of
CTZ paste; thus, it was not possible to compare the
results obtained in this study with those of other published studies.
The normal reaction to any tissue injury tends to
manifest as an acute neutrophilic inflammatory infiltrate (Serhan 2010). Given that acute histopathologic
tissue behaviour is similar across all groups, studies
that aim to test the biocompatibility of a drug that
may cause a tissue reaction should be focused on
chronic inflammatory reactions. In this study, the
pastes and the control group showed variable chronic
inflammatory processes at different experimental
times.
A histologic hallmark of successful pulp therapy is
the development of reparative tissue (Torneck 1966).
In the present study, a significant difference in the density of collagen fibres was observed between the two
pastes at the end of the experiment. The difference was
characterized by a decrease in the collagen fibres in
specimens containing either the CTZ or the calcium
hydroxide paste compared with the negative control.
This finding is important because the two pastes
behaved in manner that favours tissue repair. The
results are consistent with other studies that reported
similar methodologies (Sousa et al. 2004, Batista et al.
414
2007, Silva et al. 2010, Marques et al. 2011, GomesFilho et al. 2012).
Other materials are used as pastes for the root
canal treatment of primary teeth, including pastes
containing zinc oxide and eugenol (ZOE) with iodoform or a paste that combines iodoform and calcium
hydroxide (Ranly & Garcia-Godoy 2000, American
Academy of Pediatric Dentistry 2012). Studies with
ZOE as a root canal filling material reported a chronic
inflammatory reaction and slow resorption in relation
to the tooth (Fuks 2000, Hauman & Love 2003b).
The iodoform pastes, when combined with calcium
hydroxide, exhibit good biological response and antimicrobial properties and are more easily reabsorbed
at the periapical region (Ranly & Garcia-Godoy
2000). However, the techniques proposed for using
these materials recommend the use of instrumentation and chemicalmechanical systems for the root
canals of primary teeth (American Academy of Pediatric Dentistry 2012).
The limitations of this study are inherent to preclinical experimental studies. Although the results
presented here do not reflect a complete analysis of
the reactions that occur in the pulp, periapical or interradicular regions, the results are important for the
preliminary assessment of the irritative potential of
the tested materials and subsequent biological
responses.
Conclusion
The initial inflammatory reactions in mice induced by
CTZ and calcium hydroxide pastes regressed during the
experimental periods; in addition, these pastes resulted
in less-dense fibrous connective tissue, compared with
the results using the empty control. These reactions
support the biocompatibility of the two pastes.
Acknowledgements
This study was supported financial by a scholarship
grant from the Coordination of Improvement of
Higher Education Personnel (CAPES).
Author contributions
CCBL participated in the project design and data
collection and analysis and interpretation of data
and wrote the article. AMCJ participated in the project design and data collection and analysis and
interpretation of data and wrote the article. MSR
participated in the analysis and interpretation of

data and wrote the article. RDM participated in the
analysis of data. MSM revised the article. MDML
participated in the study preparation and revised
the article. LFADM conceived the study, participated
in the study preparation and coordination, and
wrote the article. All of the authors read and
approved the final manuscript.
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doi:10.1111/iej.

Biocompatibility of root filling pastes used in

nior2, M. S. Rizzo3, R. D. Moura4, M. S. Moura5, M. D. M.

Aim To evaluate the biocompatibility of two pastes

area of the region in direct contact with the open tube.

Keywords: biocompatibility, calcium hydroxide,

canal disinfection in primary teeth with pulp necrosis

International Endodontic Journal, 48, 405416, 2015

Materials biocompatibility Lima et al.

Dentistry 2012). The use of calcium hydroxide (Ca

Materials and methods

International Endodontic Journal, 48, 405416, 2015

University of Piau (UFPI) (opinion No. 067/12) and

Root canal pastes

Lima et al. Materials biocompatibility

pharmacy (Teresina, PI, Brazil), and the paste comprised

Experimental design for subcutaneous implantation

tubing, subcutaneous tissue and skin, was removed

International Endodontic Journal, 48, 405416, 2015

Lima et al. Materials biocompatibility

participated in the analysis and interpretation of

Hauman CHJ, Love RM (2003b) Biocompatibility of dental

International Endodontic Journal, 48, 405416, 2015

Moderately increased (Fig. 3a)

Mild collagen fibre density (Fig. 2d)

Severe; mononuclear and occasional neutrophils

Moderate; predominantly mononuclear, with the

Moderately increased (Fig. 3c)

Collagen fibres admixed with extracellular matrix

Severe; presence of macrophages, lymphocytes,

Severely increased (Fig. 3b)

Mild collagen fibre density (Fig. 2e)

Individual collagen fibres and alternating extracellular

Experimental period (days)

Inflammatory infiltrate (Fig. 4aj)

Tissue thickness (Fig. 3aj)

Calcium hydroxide paste

Formation of collagen fibres (Fig. 2aj)

Lima et al. Materials biocompatibility

International Endodontic Journal, 48, 405416, 2015

Materials biocompatibility Lima et al.

International Endodontic Journal, 48, 405416, 2015

able, for primary teeth with necrotic pulps (Silva et al.

Lima et al. Materials biocompatibility

International Endodontic Journal, 48, 405416, 2015

Materials biocompatibility Lima et al.

International Endodontic Journal, 48, 405416, 2015

Lima et al. Materials biocompatibility

M = mean; SD = standard deviation.

young children with behavioural problems. Another

Compatibility with living tissue in the periapical or

International Endodontic Journal, 48, 405416, 2015

Materials biocompatibility Lima et al.

used in dentistry due to its biocompatible properties

International Endodontic Journal, 48, 405416, 2015

Lima et al. Materials biocompatibility

participated in the analysis and interpretation of

Hauman CHJ, Love RM (2003b) Biocompatibility of dental

International Endodontic Journal, 48, 405416, 2015

Materials biocompatibility Lima et al.

International Endodontic Journal, 48, 405416, 2015

of the biocompatibility of materials used in apical surgery.

Das könnte Ihnen auch gefallen

M = mean; SD = standard deviation.